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PCR_Testing_of_colonies - PCR Testing of Plate Colonies...

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PCR Testing of Plate Colonies April 6, 2009 When you have a plate with many colonies on it and you need to test many or all of them for the presence of a specific piece of DNA (such as a specific insert in a plasmid) it can be tedious and time consuming to do minipreps for them all. This is a less labor-intensive method of checking the colonies, provided you have primers to amplify the piece of DNA you are searching for. General procedure: 1. Warm up and mark an LB-amp plate and place 10 µl of TE to a separate microfuge tube for each colony to be picked. The LB-amp plate will eventually represent a master plate for each of your colonies. 2. Pick colonies with a toothpick and gently streak the each toothpick onto a localized, marked area of the LB-amp plate. This will be your stock colony plate. Be sure you know which colony represents which toothpick. After picking all your colonies, place the master plate at 37°C overnight.
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